Skip to main content
PMC home page
Genetics logoLink to Genetics
. 2001 Oct;159(2):635–645. doi: 10.1093/genetics/159.2.635

Interallelic complementation at the Drosophila melanogaster gastrulation defective locus defines discrete functional domains of the protein.

G Ponomareff 1, H Giordano 1, Y DeLotto 1, R DeLotto 1
PMCID: PMC1461824  PMID: 11606540

Abstract

The gastrulation defective (gd) locus encodes a novel serine protease that is involved in specifying the dorsal-ventral axis during embryonic development. Mutant alleles of gd have been classified into three complementation groups, two of which exhibit strong interallelic (intragenic) complementation. To understand the molecular basis of this interallelic complementation, we examined the complementation behavior of additional mutant alleles and sequenced alleles in all complementation groups. The data suggest that there are two discrete functional domains of Gd. A two-domain model of Gd suggesting that it is structurally similar to mammalian complement factors C2 and B has been previously proposed. To test this model we performed SP6 RNA microinjection to assay for activities associated with various domains of Gd. The microinjection data are consistent with the complement factor C2/B-like model. Site-directed mutagenesis suggests that Gd functions as a serine protease. An allele-specific interaction between an autoactivating form of Snake (Snk) and a gd allele altered in the protease domain suggests that Gd directly activates Snk in a protease activation cascade. We propose a model in which Gd is expressed during late oogenesis and bound within the perivitelline space but only becomes catalytically active during embryogenesis.

Full Text

The Full Text of this article is available as a PDF (250.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arlaud G. J., Volanakis J. E., Thielens N. M., Narayana S. V., Rossi V., Xu Y. The atypical serine proteases of the complement system. Adv Immunol. 1998;69:249–307. [PubMed] [Google Scholar]
  2. Bier E., Jan L. Y., Jan Y. N. rhomboid, a gene required for dorsoventral axis establishment and peripheral nervous system development in Drosophila melanogaster. Genes Dev. 1990 Feb;4(2):190–203. doi: 10.1101/gad.4.2.190. [DOI] [PubMed] [Google Scholar]
  3. Chasan R., Anderson K. V. The role of easter, an apparent serine protease, in organizing the dorsal-ventral pattern of the Drosophila embryo. Cell. 1989 Feb 10;56(3):391–400. doi: 10.1016/0092-8674(89)90242-0. [DOI] [PubMed] [Google Scholar]
  4. DeLotto R. Gastrulation defective, a complement factor C2/B-like protease, interprets a ventral prepattern in Drosophila. EMBO Rep. 2001 Aug;2(8):721–726. doi: 10.1093/embo-reports/kve153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DeLotto R., Spierer P. A gene required for the specification of dorsal-ventral pattern in Drosophila appears to encode a serine protease. Nature. 1986 Oct 23;323(6090):688–692. doi: 10.1038/323688a0. [DOI] [PubMed] [Google Scholar]
  6. DeLotto Y., Smith C., DeLotto R. Multiple isoforms of the Drosophila Spätzle protein are encoded by alternatively spliced maternal mRNAs in the precellular blastoderm embryo. Mol Gen Genet. 2001 Jan;264(5):643–652. doi: 10.1007/s004380000350. [DOI] [PubMed] [Google Scholar]
  7. Dissing M., Giordano H., DeLotto R. Autoproteolysis and feedback in a protease cascade directing Drosophila dorsal-ventral cell fate. EMBO J. 2001 May 15;20(10):2387–2393. doi: 10.1093/emboj/20.10.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Han J. H., Lee S. H., Tan Y. Q., LeMosy E. K., Hashimoto C. Gastrulation defective is a serine protease involved in activating the receptor toll to polarize the Drosophila embryo. Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):9093–9097. doi: 10.1073/pnas.97.16.9093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Konrad K. D., Goralski T. J., Mahowald A. P. Developmental genetics of the gastrulation defective locus in Drosophila melanogaster. Dev Biol. 1988 May;127(1):133–142. doi: 10.1016/0012-1606(88)90195-9. [DOI] [PubMed] [Google Scholar]
  10. Konrad K. D., Goralski T. J., Mahowald A. P., Marsh J. L. The gastrulation defective gene of Drosophila melanogaster is a member of the serine protease superfamily. Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6819–6824. doi: 10.1073/pnas.95.12.6819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LeMosy E. K., Tan Y. Q., Hashimoto C. Activation of a protease cascade involved in patterning the Drosophila embryo. Proc Natl Acad Sci U S A. 2001 Apr 10;98(9):5055–5060. doi: 10.1073/pnas.081026598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mann K. G., Jenny R. J., Krishnaswamy S. Cofactor proteins in the assembly and expression of blood clotting enzyme complexes. Annu Rev Biochem. 1988;57:915–956. doi: 10.1146/annurev.bi.57.070188.004411. [DOI] [PubMed] [Google Scholar]
  13. Mohler J. D. Developmental genetics of the Drosophila egg. I. Identification of 59 sex-linked cistrons with maternal effects on embryonic development. Genetics. 1977 Feb;85(2):259–272. doi: 10.1093/genetics/85.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Morisato D., Anderson K. V. Signaling pathways that establish the dorsal-ventral pattern of the Drosophila embryo. Annu Rev Genet. 1995;29:371–399. doi: 10.1146/annurev.ge.29.120195.002103. [DOI] [PubMed] [Google Scholar]
  15. Morisato D., Anderson K. V. The spätzle gene encodes a component of the extracellular signaling pathway establishing the dorsal-ventral pattern of the Drosophila embryo. Cell. 1994 Feb 25;76(4):677–688. doi: 10.1016/0092-8674(94)90507-x. [DOI] [PubMed] [Google Scholar]
  16. Neuman-Silberberg F. S., Schüpbach T. The Drosophila dorsoventral patterning gene gurken produces a dorsally localized RNA and encodes a TGF alpha-like protein. Cell. 1993 Oct 8;75(1):165–174. [PubMed] [Google Scholar]
  17. Oglesby T. J., Accavitti M. A., Volanakis J. E. Evidence for a C4b binding site on the C2b domain of C2. J Immunol. 1988 Aug 1;141(3):926–931. [PubMed] [Google Scholar]
  18. Roth S. Axis determination. Proteolytic generation of a morphogen. Curr Biol. 1994 Aug 1;4(8):755–757. doi: 10.1016/s0960-9822(00)00170-6. [DOI] [PubMed] [Google Scholar]
  19. Roth S., Neuman-Silberberg F. S., Barcelo G., Schüpbach T. cornichon and the EGF receptor signaling process are necessary for both anterior-posterior and dorsal-ventral pattern formation in Drosophila. Cell. 1995 Jun 16;81(6):967–978. doi: 10.1016/0092-8674(95)90016-0. [DOI] [PubMed] [Google Scholar]
  20. Roth S., Schüpbach T. The relationship between ovarian and embryonic dorsoventral patterning in Drosophila. Development. 1994 Aug;120(8):2245–2257. doi: 10.1242/dev.120.8.2245. [DOI] [PubMed] [Google Scholar]
  21. Roth S., Stein D., Nüsslein-Volhard C. A gradient of nuclear localization of the dorsal protein determines dorsoventral pattern in the Drosophila embryo. Cell. 1989 Dec 22;59(6):1189–1202. doi: 10.1016/0092-8674(89)90774-5. [DOI] [PubMed] [Google Scholar]
  22. Rushlow C. A., Han K., Manley J. L., Levine M. The graded distribution of the dorsal morphogen is initiated by selective nuclear transport in Drosophila. Cell. 1989 Dec 22;59(6):1165–1177. doi: 10.1016/0092-8674(89)90772-1. [DOI] [PubMed] [Google Scholar]
  23. Schüpbach T. Germ line and soma cooperate during oogenesis to establish the dorsoventral pattern of egg shell and embryo in Drosophila melanogaster. Cell. 1987 Jun 5;49(5):699–707. doi: 10.1016/0092-8674(87)90546-0. [DOI] [PubMed] [Google Scholar]
  24. Stein D., Nüsslein-Volhard C. Multiple extracellular activities in Drosophila egg perivitelline fluid are required for establishment of embryonic dorsal-ventral polarity. Cell. 1992 Feb 7;68(3):429–440. doi: 10.1016/0092-8674(92)90181-b. [DOI] [PubMed] [Google Scholar]
  25. Stein D., Roth S., Vogelsang E., Nüsslein-Volhard C. The polarity of the dorsoventral axis in the Drosophila embryo is defined by an extracellular signal. Cell. 1991 May 31;65(5):725–735. doi: 10.1016/0092-8674(91)90381-8. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES